scholarly journals Kilohertz two-photon fluorescence microscopy imaging of neural activity in vivo

2019 ◽  
Author(s):  
Jianglai Wu ◽  
Yajie Liang ◽  
Shuo Chen ◽  
Ching-Lung Hsu ◽  
Mariya Chavarha ◽  
...  
Keyword(s):  
Neural Activity ◽  
Laser Scanning ◽  
Imaging Method ◽  
Spatiotemporal Resolution ◽  
Microscopy Imaging ◽  
Two Photon ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence ◽  
The Brain

Understanding information processing in the brain requires us to monitor neural activity in vivo at high spatiotemporal resolution. Using an ultrafast two-photon fluorescence microscope (2PFM) empowered by all-optical laser scanning, we imaged neural activity in vivo at up to 3,000 frames per second and submicron spatial resolution. This ultrafast imaging method enabled monitoring of both supra- and sub-threshold electrical activity down to 345 μm below the brain surface in head fixed awake mice.

scholarly journals Kilohertz two-photon fluorescence microscopy imaging of neural activity in vivo

2020 ◽  
Vol 17 (3) ◽  
pp. 287-290 ◽  
Author(s):  
Jianglai Wu ◽  
Yajie Liang ◽  
Shuo Chen ◽  
Ching-Lung Hsu ◽  
Mariya Chavarha ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Neural Activity ◽  
Microscopy Imaging ◽  
Two Photon ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

scholarly journals High-speed volumetric two-photon fluorescence imaging of neurovascular dynamics

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiang Lan Fan ◽  
Jose A. Rivera ◽  
Wei Sun ◽  
John Peterson ◽  
Henry Haeberle ◽  
...  
Keyword(s):  
Blood Flow ◽  
High Speed ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Laser Scanning Microscopy ◽  
Fluorescence Signal ◽  
Imaging Method ◽  
Spatiotemporal Resolution ◽  
Two Photon

AbstractUnderstanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.

scholarly journals In Vivo Mammalian Brain Imaging Using One- and Two-Photon Fluorescence Microendoscopy

2004 ◽  
Vol 92 (5) ◽  
pp. 3121-3133 ◽  
Author(s):  
Juergen C. Jung ◽  
Amit D. Mehta ◽  
Emre Aksay ◽  
Raymond Stepnoski ◽  
Mark J. Schnitzer
Keyword(s):  
Laser Scanning ◽  
Frame Rate ◽  
Mammalian Brain ◽  
Cell Dynamics ◽  
Ca1 Neurons ◽  
Two Photon ◽  
Two Photon Fluorescence ◽  
Rats And Mice ◽  
Photon Fluorescence

One of the major limitations in the current set of techniques available to neuroscientists is a dearth of methods for imaging individual cells deep within the brains of live animals. To overcome this limitation, we developed two forms of minimally invasive fluorescence microendoscopy and tested their abilities to image cells in vivo. Both one- and two-photon fluorescence microendoscopy are based on compound gradient refractive index (GRIN) lenses that are 350–1,000 μm in diameter and provide micron-scale resolution. One-photon microendoscopy allows full-frame images to be viewed by eye or with a camera, and is well suited to fast frame-rate imaging. Two-photon microendoscopy is a laser-scanning modality that provides optical sectioning deep within tissue. Using in vivo microendoscopy we acquired video-rate movies of thalamic and CA1 hippocampal red blood cell dynamics and still-frame images of CA1 neurons and dendrites in anesthetized rats and mice. Microendoscopy will help meet the growing demand for in vivo cellular imaging created by the rapid emergence of new synthetic and genetically encoded fluorophores that can be used to label specific brain areas or cell classes.

scholarly journals Aggregation-induced emission nanoparticles for in vivo three-photon fluorescence microscopic rat brain angiography

2019 ◽  
Vol 12 (06) ◽  
pp. 1950012 ◽  
Author(s):  
Hequn Zhang ◽  
Weisi Xie ◽  
Ming Chen ◽  
Liang Zhu ◽  
Zhe Feng ◽  
...  
Keyword(s):  
Rat Brain ◽  
Blood Vessels ◽  
Laser Scanning ◽  
Near Infrared ◽  
Brain Disease ◽  
Laser Scanning Microscopy ◽  
High Signal ◽  
Imaging Method ◽  
Photon Fluorescence

Rodents are popular biological models for physiological and behavioral research in neuroscience and rats are better models than mice due to their higher genome similarity to human and more accessible surgical procedures. However, rat brain is larger than mice brain and it needs powerful imaging tools to implement better penetration against the scattering of the thicker brain tissue. Three-photon fluorescence microscopy (3PFM) combined with near-infrared (NIR) excitation has great potentials for brain circuits imaging because of its abilities of anti-scattering, deep-tissue imaging, and high signal-to-noise ratio (SNR). In this work, a type of AIE luminogen with red fluorescence was synthesized and encapsulated with Pluronic F-127 to make up form nanoparticles (NPs). Bright DCDPP-2TPA NPs were employed for in vivo three-photon fluorescent laser scanning microscopy of blood vessels in rats brain under 1550[Formula: see text]nm femtosecond laser excitation. A fine three-dimensional (3D) reconstruction up to the deepness of 600[Formula: see text][Formula: see text]m was achieved and the blood flow velocity of a selected vessel was measured in vivo as well. Our 3PFM deep brain imaging method simultaneously recorded the morphology and function of the brain blood vessels in vivo in the rat model. Using this angiography combined with the arsenal of rodent’s brain disease, models can accelerate the neuroscience research and clinical diagnosis of brain disease in the future.

Silole-Based Red Fluorescent Organic Dots for Bright Two-Photon Fluorescence In vitro Cell and In vivo Blood Vessel Imaging

Small ◽  
2015 ◽  
Vol 12 (6) ◽  
pp. 782-792 ◽  
Author(s):  
Bin Chen ◽  
Guangxue Feng ◽  
Bairong He ◽  
Chiching Goh ◽  
Shidang Xu ◽  
...  
Keyword(s):  
Blood Vessel ◽  
Two Photon ◽  
Vessel Imaging ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

scholarly journals Two-Photon Microscopy Allows Imaging and Characterization of Cochlear Microvasculature In Vivo

2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Friedrich Ihler ◽  
Mattis Bertlich ◽  
Bernhard Weiss ◽  
Steffen Dietzel ◽  
Martin Canis
Keyword(s):  
Fluorescence Microscopy ◽  
Inner Ear ◽  
Ex Vivo ◽  
Small Scale ◽  
Published Data ◽  
Cochlear Blood Flow ◽  
Two Photon ◽  
Two Photon Fluorescence ◽  
Photon Fluorescence

Impairment of cochlear blood flow has been discussed as factor in the pathophysiology of various inner ear disorders. However, the microscopic study of cochlear microcirculation is limited due to small scale and anatomical constraints. Here, two-photon fluorescence microscopy is applied to visualize cochlear microvessels. Guinea pigs were injected with Fluorescein isothiocyanate- or Texas red-dextrane as plasma marker. Intravital microscopy was performed in four animals and explanted cochleae from four animals were studied. The vascular architecture of the cochlea was visualized up to a depth of90.0±22.7 μm. Imaging yielded a mean contrast-to-noise ratio (CNR) of3.3±1.7. Mean diameter in vivo was16.5±6.0 μm for arterioles and8.0±2.4 μm for capillaries. In explanted cochleae, the diameter of radiating arterioles and capillaries was measured with12.2±1.6 μm and6.6±1.0 μm, respectively. The difference between capillaries and arterioles was statistically significant in both experimental setups (P<0.001andP=0.022, two-way ANOVA). Measured vessel diameters in vivo and ex vivo were in agreement with published data. We conclude that two-photon fluorescence microscopy allows the investigation of cochlear microvessels and is potentially a valuable tool for inner ear research.

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